Dendrobium huoshanense protocorm functional food with enhanced immunity

Abstract

The application discloses a Dendrobium huoshanense protocorm functional food with an enhanced immunity effect, which is prepared by compounding Dendrobium huoshanense protocorm extract, double-spore mushroom extract and Raphiolepis umbellate leaf extract according to a specific weight ratio. The Dendrobium huoshanense protocorm functional food with the enhanced immunity effect prepared by the application can significantly enhance the cell immunity function, the humoral immunity function, the monocyte-macrophage function and the NK cell activity of test animals through animal test verification.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of functional food development, and particularly relates to a Dendrobium huoshanense protocorm functional food with the function of enhancing immunity. BACKGROUND

[0002] At present, the products processed from the medicinal and edible raw materials such as Dendrobium huoshanense, Agaricus bisporus and Radermachera sinensis have the problems of low value of functional factor extraction, uncertain efficacy and low-end products, therefore, it has become the development trend of the medicinal and edible raw material industry to establish the high value extraction of functional factors of medicinal and edible raw materials, the accurate evaluation technology of efficacy, and the key technology system of creating high-end health products based on the above, and to develop high-end health products.

[0003] Dendrobium huoshanense, Agaricus bisporus and Radermachera sinensis are all characteristic medicinal and edible raw materials, and have a long history of medical use and great market potential, but at present, the processed products are mainly low-end primary processing products, and there are few deep processing products, which greatly limits the development of the industry. In order to solve the problem of low value of functional factor extraction of the medicinal and edible raw materials such as Dendrobium huoshanense, Agaricus bisporus and Radermachera sinensis in the health industry and promote the development of the industry, the present application develops a Dendrobium functional food with the function of enhancing immunity. SUMMARY

[0004] The present application discloses a Dendrobium huoshanense protocorm functional food with the function of enhancing immunity, which is composed of 17-19 parts by weight of Dendrobium huoshanense protocorm extract, 11-13 parts by weight of Agaricus bisporus extract and 7-9 parts by weight of Radermachera sinensis leaf extract. The Dendrobium huoshanense protocorm functional food with the function of enhancing immunity prepared by the present application can significantly enhance the cellular immune function, humoral immune function, monocyte-macrophage function and NK cell activity of the test animals through animal test verification.

[0005] The specific implementation steps of the present application are as follows:

[0006] 1. Preparation of Dendrobium huoshanense protocorm extract

[0007] (1) Preparation of Dendrobium huoshanense protocorm alcohol extract: take Dendrobium huoshanense protocorm ultrafine powder as raw material, and anhydrous ethanol as extraction agent, under the conditions of vacuum degree 0.1 Mpa, ultrasonic power 300 W, temperature 30℃ and solid-liquid ratio 1:30, extract for 30 min under the condition of vacuum and ultrasonic extraction, and then concentrate and dry the extract to obtain Dendrobium huoshanense protocorm alcohol extract;

[0008] (2) Preparation of the water extract of D. candidum protocorm: The defatted D. candidum protocorm ultrafine powder was used as raw material, and pure water was used as the extraction agent. The vacuum degree was 0.1 Mpa, the ultrasonic power was 300 W, the temperature was 60℃, the solid-liquid ratio was 1:30, and the vacuum ultrasonic extraction was performed for 30 min. The water extract of D. candidum protocorm was obtained by concentrating and drying the extraction liquid.

[0009] (3) Preparation of the extract of D. candidum protocorm: The alcohol extract and the water extract of D. candidum protocorm were mixed in a proportion according to the extraction rate to obtain the extract of D. candidum protocorm.

[0010] 2. Preparation of the extract of Agaricus bisporus

[0011] (1) Preparation of the alcohol extract of Agaricus bisporus: The Agaricus bisporus ultrafine powder was used as raw material, and anhydrous ethanol was used as the extraction agent. The vacuum degree was 0.1 Mpa, the ultrasonic power was 300 W, the temperature was 30℃, the solid-liquid ratio was 1:30, and the vacuum ultrasonic extraction was performed for 30 min. The alcohol extract of Agaricus bisporus was obtained by concentrating and drying the extraction liquid.

[0012] (2) Preparation of the water extract of Agaricus bisporus: The defatted Agaricus bisporus ultrafine powder was used as raw material, and pure water was used as the extraction agent. The vacuum degree was 0.1 Mpa, the ultrasonic power was 300 W, the temperature was 70℃, the solid-liquid ratio was 1:30, and the vacuum ultrasonic extraction was performed for 30 min. The water extract of Agaricus bisporus was obtained by concentrating and drying the extraction liquid.

[0013] (3) Preparation of the extract of Agaricus bisporus: The alcohol extract and the water extract of Agaricus bisporus were mixed in a proportion according to the extraction rate to obtain the extract of Agaricus bisporus.

[0014] 3. Preparation of the extract of Premna microphylla Turcz.

[0015] (1) Preparation of the alcohol extract of Premna microphylla Turcz. leaves: The Premna microphylla Turcz. leaf ultrafine powder was used as raw material, and anhydrous ethanol was used as the extraction agent. The vacuum degree was 0.1 Mpa, the ultrasonic power was 300 W, the temperature was 30℃, the solid-liquid ratio was 1:30, and the vacuum ultrasonic extraction was performed for 30 min. The alcohol extract of Premna microphylla Turcz. leaves was obtained by concentrating and drying the extraction liquid.

[0016] (2) Preparation of the water extract of Premna microphylla Turcz. leaves: The defatted Premna microphylla Turcz. leaf ultrafine powder was used as raw material, and pure water was used as the extraction agent. The vacuum degree was 0.1 Mpa, the ultrasonic power was 300 W, the temperature was 80℃, the solid-liquid ratio was 1:30, and the vacuum ultrasonic extraction was performed for 30 min. The water extract of Premna microphylla Turcz. leaves was obtained by concentrating and drying the extraction liquid.

[0017] (3) Preparation of the extract of Premna microphylla Turcz. leaves: The alcohol extract and the water extract of Premna microphylla Turcz. leaves were mixed in a proportion according to the extraction rate to obtain the extract of Premna microphylla Turcz. leaves.

[0018] 4. Preparation of the functional food of D. candidum protocorm with the function of enhancing immunity

[0019] 17-19 parts by weight of Dendrobium candidum protocorm extract, 11-13 parts by weight of Agaricus bisporus extract, and 7-9 parts by weight of Premna microphylla Turcz leaf extract are weighed and uniformly mixed to prepare a Dendrobium functional food with the effect of enhancing immunity.

[0020] The beneficial technical effects of the present application are embodied in:

[0021] (1) The alcohol-soluble components and water-soluble components in the Dendrobium candidum protocorm, Agaricus bisporus, and Premna microphylla Turcz leaves are extracted separately, and then the alcohol extract and water extract of the Dendrobium candidum protocorm, Agaricus bisporus, and Premna microphylla Turcz leaves are mixed in equal proportions according to the extraction rate. Compared with directly consuming the Dendrobium candidum protocorm, Agaricus bisporus, and Premna microphylla Turcz leaves, the functional component utilization rate of the extract is high, and the effect is significant.

[0022] (2) The Dendrobium candidum protocorm extract, Agaricus bisporus extract, and Premna microphylla Turcz leaf extract are combined in a specific weight ratio. Through experimental verification, the combined Dendrobium candidum protocorm extract, Agaricus bisporus extract, and Premna microphylla Turcz leaf extract has a synergistic effect on enhancing the cell immune function, humoral immune function, monocyte-macrophage function, and NK cell activity of test animals, and has a significant immune-enhancing function. DETAILED DESCRIPTION

[0023] The present application will be further described below in conjunction with specific examples. The features and advantages of the present application will become more apparent as the description proceeds, but the exemplary examples are only used to illustrate the present application and do not constitute any limitation on the scope of the present application.

[0024] Example 1

[0025] 1. Preparation of Dendrobium candidum protocorm extract

[0026] (1) Preparation of Dendrobium candidum protocorm alcohol extract: Dendrobium candidum protocorm ultrafine powder is used as the raw material, and anhydrous ethanol is used as the extraction agent. Under the conditions of a vacuum degree of 0.1 Mpa, an ultrasonic power of 300 W, a temperature of 30℃, and a solid-liquid ratio of 1:30, the Dendrobium candidum protocorm alcohol extract is prepared by vacuum and ultrasonic extraction for 30 min. The extract is concentrated and dried to obtain the Dendrobium candidum protocorm alcohol extract.

[0027] (2) Preparation of Dendrobium candidum protocorm water extract: defatted Dendrobium candidum protocorm ultrafine powder is used as the raw material, and pure water is used as the extraction agent. Under the conditions of a vacuum degree of 0.1 Mpa, an ultrasonic power of 300 W, a temperature of 60℃, and a solid-liquid ratio of 1:30, the Dendrobium candidum protocorm water extract is prepared by vacuum and ultrasonic extraction for 30 min. The extract is concentrated and dried to obtain the Dendrobium candidum protocorm water extract.

[0028] (3) Preparation of Dendrobium huoshanense protocorm extract: the alcohol extract and the water extract of Dendrobium huoshanense protocorm are mixed in equal proportion according to the extraction rate to prepare the Dendrobium huoshanense protocorm extract.

[0029] 2. Preparation of Agaricus bisporus extract

[0030] (1) Preparation of Agaricus bisporus alcohol extract: under the conditions of vacuum degree 0.1 Mpa, ultrasonic power 300 W, temperature 30℃, and solid-liquid ratio 1:30, Agaricus bisporus ultrafine powder is extracted by vacuum and ultrasonic extraction for 30 min with anhydrous ethanol as the extracting agent, and the extract is concentrated and dried to obtain the Agaricus bisporus alcohol extract;

[0031] (2) Preparation of Agaricus bisporus water extract: under the conditions of vacuum degree 0.1 Mpa, ultrasonic power 300 W, temperature 70℃, and solid-liquid ratio 1:30, defatted Agaricus bisporus ultrafine powder is extracted by vacuum and ultrasonic extraction for 30 min with pure water as the extracting agent, and the extract is concentrated and dried to obtain the Agaricus bisporus water extract;

[0032] (3) Preparation of Agaricus bisporus extract: the Agaricus bisporus alcohol extract and the water extract are mixed in equal proportion according to the extraction rate to prepare the Agaricus bisporus extract.

[0033] 3. Preparation of Premna microphylla Turcz. leaf extract

[0034] (1) Preparation of Premna microphylla Turcz. leaf alcohol extract: under the conditions of vacuum degree 0.1 Mpa, ultrasonic power 300 W, temperature 30℃, and solid-liquid ratio 1:30, Premna microphylla Turcz. leaf ultrafine powder is extracted by vacuum and ultrasonic extraction for 30 min with anhydrous ethanol as the extracting agent, and the extract is concentrated and dried to obtain the Premna microphylla Turcz. leaf alcohol extract;

[0035] (2) Preparation of Premna microphylla Turcz. leaf water extract: under the conditions of vacuum degree 0.1 Mpa, ultrasonic power 300 W, temperature 80℃, and solid-liquid ratio 1:30, defatted Premna microphylla Turcz. leaf ultrafine powder is extracted by vacuum and ultrasonic extraction for 30 min with pure water as the extracting agent, and the extract is concentrated and dried to obtain the Premna microphylla Turcz. leaf water extract;

[0036] (3) Preparation of Premna microphylla Turcz. leaf extract: the Premna microphylla Turcz. leaf alcohol extract and the water extract are mixed in equal proportion according to the extraction rate to prepare the Premna microphylla Turcz. leaf extract.

[0037] 4. Preparation of Dendrobium huoshanense protocorm functional food with the function of enhancing immunity

[0038] 19 parts by weight of Dendrobium huoshanense protocorm extract, 13 parts by weight of Agaricus bisporus extract, and 9 parts by weight of Premna microphylla Turcz. leaf extract are weighed and uniformly mixed to prepare the Dendrobium huoshanense protocorm functional food with the function of enhancing immunity.

[0039] 5. Determination of the function of enhancing immunity

[0040] (1) Test grouping: The immunity-enhancing function test uses male Kunming mice as test animals, and the test is divided into blank control group, Dendrobium huoshanense protocorm extract group, Agaricus bisporus extract group, Premna microphylla Turcz. leaf extract group, and extract combination group. The test indicators include mouse spleen lymphocyte transformation test, delayed-type hypersensitivity test, antibody-producing cell detection, serum hemolysin determination, mouse carbon clearance test, mouse peritoneal macrophage phagocytosis of chicken red blood cells test, and NK cell activity determination.

[0041] (2) Test method: Refer to the immunity-enhancing test method in "Health Food Function Test and Evaluation Method". The blank control group is given distilled water by gavage, the Dendrobium huoshanense protocorm extract group, Agaricus bisporus extract group, Premna microphylla Turcz. leaf extract group, and extract combination group are given Dendrobium huoshanense protocorm extract, Agaricus bisporus extract, Premna microphylla Turcz. leaf extract, and extract combination by gavage at a dosage of 600 mg / kg / d, respectively. The gavage volume is 0.2 mL, and the gavage time is 30 days.

[0042] (3) Test results

[0043] Table 1: Test results of mouse spleen lymphocyte transformation test

[0044] Group Number of animals Lymphocyte proliferation ability (OD difference) Blank control group 15 0.029 ± 0.013 b ]] Dendrobium huoshanense protocorm extract group 15 0.051 ± 0.019 ab ]] Agaricus bisporus extract group 15 0.045 ± 0.021 ab ]] Radermachera sinica leaf extract group 15 0.033 ± 0.017 b ]] Extract combination group 15 0.071 ± 0.023 a ]]

[0045] Note: a Significant difference compared with the blank control group, b Significant difference compared with the extract combination group, P<0.05.

[0046] From the test results of mouse spleen lymphocyte transformation test in Table 1, when the three extracts are used alone, the test data of the Dendrobium huoshanense protocorm extract group and the Agaricus bisporus extract group are higher than those of the blank control group and lower than those of the extract combination group. The test data of the Premna microphylla Turcz. leaf extract group is not different from that of the blank control group. The test data of the extract combination group is not only significantly (P<0.05) higher than that of the blank control group, but also significantly (P<0.05) higher than that of each extract alone group, indicating that the combination of Dendrobium huoshanense protocorm extract, Agaricus bisporus extract, and Premna microphylla Turcz. leaf extract has a synergistic effect on increasing CoA-induced mouse spleen lymphocyte transformation.

[0047] Table 2: Test results of delayed-type hypersensitivity test

[0048] Group Number of animals Paw thickness difference (mm) Blank control group 15 0.41 ± 0.12 b ]] Dendrobium huoshanense protocorm extract group 15 0.39 ± 0.19 b ]] Agaricus bisporus extract group 15 0.42 ± 0.13 b ]] Radermachera sinica leaf extract group 15 0.53 ± 0.17 ab ]] Extract combination group 15 0.71 ± 0.23 a ]]

[0049] Note: a Significant difference compared with the blank control group, b Significant difference compared with the extract combination group, P<0.05.

[0050] From the results of the delayed allergy experiment in Table 2, when the three extracts were used alone, the determination data of the D. candidum protocorm extract group and the double-spore mushroom extract group were not different from the blank control group, the determination data of the R. javanicum leaf extract group was higher than the blank control group and lower than the extract combination group. The determination data of the extract combination group was not only significantly (P<0.05) higher than the blank control group, but also significantly (P<0.05) higher than each of the extract alone groups, indicating that the D. candidum protocorm extract, double-spore mushroom extract and R. javanicum leaf extract combined together have a synergistic effect of improving the delayed allergy.

[0051] Table 3 Determination results of antibody-producing cell detection experiment

[0052] Group Number of animals Number of hemolytic plaques (x 10 3 / total spleen Blank control group 15 39.18 ± 9.61 b ]] Dendrobium huoshanense protocorm extract group 15 51.39 ± 9.19 ab ]] Agaricus bisporus extract group 15 53.42 ± 7.13 ab ]] Radermachera sinica leaf extract group 15 52.53 ± 8.17 ab ]] Extract combination group 15 81.71 ± 7.23 a ]]

[0053] Note: a Significant difference compared with the blank control group, b Significant difference compared with the extract combination group, P<0.05.

[0054] From the determination results of the antibody-producing cell detection experiment in Table 3, when the three extracts were used alone, the determination data of the D. candidum protocorm extract group, double-spore mushroom extract group and R. javanicum leaf extract group were all higher than the blank control group and lower than the extract combination group. The determination data of the extract combination group was not only significantly (P<0.05) higher than the blank control group, but also significantly (P<0.05) higher than each of the extract alone groups, indicating that the D. candidum protocorm extract, double-spore mushroom extract and R. javanicum leaf extract combined together have a synergistic effect of increasing the number of antibody-producing cells.

[0055] Table 4 Determination results of serum hemolysin determination experiment

[0056] Group Number of animals Antibody volume Blank control group 15 137.71 ± 11.33 b ]] Dendrobium huoshanense protocorm extract group 15 162.67 ± 9.89 ab ]] Agaricus bisporus extract group 15 163.23 ± 8.34 ab ]] Radermachera sinica leaf extract group 15 162.01 ± 9.09 ab ]] Extract combination group 15 189.93 ± 10.07 a ]]

[0057] Note: a Significant difference compared with the blank control group, b Significant difference compared with the extract combination group, P<0.05.

[0058] From the determination results of the serum hemolysin determination experiment in Table 4, when the three extracts were used alone, the determination data of the D. candidum protocorm extract group, double-spore mushroom extract group and R. javanicum leaf extract group were all higher than the blank control group and lower than the extract combination group. The determination data of the extract combination group was not only significantly (P<0.05) higher than the blank control group, but also significantly (P<0.05) higher than each of the extract alone groups, indicating that the D. candidum protocorm extract, double-spore mushroom extract and R. javanicum leaf extract combined together have a synergistic effect of improving the serum hemolysin level.

[0059] Table 5 Determination results of carbon clearance experiment

[0060] Group Number of animals Phagocytic index Blank control group 15 3.71 ± 0.43 b ]] Dendrobium huoshanense protocorm extract group 15 5.06 ± 0.91 ab ]] Agaricus bisporus extract group 15 4.01 ± 0.45 b ]] Radermachera sinica leaf extract group 15 5.11 ± 0.19 ab ]] Extract combination group 15 7.63 ± 0.77 a ]]

[0061] Note: a Significant difference compared with the blank control group, b Significant difference compared with the extract combination group, P<0.05.

[0062] From the determination results of carbon clearance experiment in Table 5, when the three extracts act alone, the determination data of the D. candidum protocorm extract group and the R. lanceolata leaf extract group are higher than that of the blank control group and lower than that of the extract combination group, and the determination data of the A. bisporus extract group has no difference with that of the blank control group. The determination data of the extract combination group is not only significantly (P<0.05) higher than that of the blank control group, but also significantly (P<0.05) higher than that of each experimental group in which the extracts act alone, indicating that the D. candidum protocorm extract, the A. bisporus extract and the R. lanceolata leaf extract combined together have the synergistic effect of improving the carbon clearance ability of mononuclear macrophages.

[0063] Table 6 Determination results of mouse peritoneal macrophage phagocytosis of chicken red blood cells experiment

[0064] Group Number of animals Chicken red blood cell phagocytic index Blank control group 15 0.27 ± 0.13 b ]] Dendrobium huoshanense protocorm extract group 15 0.63 ± 0.21 ab ]] Agaricus bisporus extract group 15 0.31 ± 0.16 b ]] Radermachera sinica leaf extract group 15 0.66 ± 0.18 ab ]] Extract combination group 15 0.93 ± 0.25 a ]]

[0065] Note: a Significant difference compared with the blank control group, b Significant difference compared with the extract combination group, P<0.05.

[0066] From the determination results of mouse peritoneal macrophage phagocytosis of chicken red blood cells experiment in Table 6, when the three extracts act alone, the determination data of the D. candidum protocorm extract group and the R. lanceolata leaf extract group are higher than that of the blank control group and lower than that of the extract combination group, and the determination data of the A. bisporus extract group has no difference with that of the blank control group. The determination data of the extract combination group is not only significantly (P<0.05) higher than that of the blank control group, but also significantly (P<0.05) higher than that of each experimental group in which the extracts act alone, indicating that the D. candidum protocorm extract, the A. bisporus extract and the R. lanceolata leaf extract combined together have the synergistic effect of improving the phagocytosis ability of mouse peritoneal macrophages on chicken red blood cells.

[0067] Table 7 Determination results of NK cell activity determination experiment

[0068] Group Number of animals NK cell activity (%) Blank control group 15 23.44 ± 0.17 b ]] Dendrobium huoshanense protocorm extract group 15 24.43 ± 0.19 b ]] Agaricus bisporus extract group 15 35.59 ± 0.09 ab ]] Radermachera sinica leaf extract group 15 23.67 ± 0.17 b ]] Extract combination group 15 49.81 ± 0.15 a ]]

[0069] Note: a Significant difference compared with the blank control group, b Significant difference compared with the extract combination group, P<0.05.

[0070] From the determination results of the NK cell activity determination experiment of Table 7, when the three extracts were used alone, the determination data of the Agaricus bisporus extract group was higher than that of the blank control group and lower than that of the extract combination group, and the determination data of the Dendrobium huoshanense protocorm extract group and the Radermachera xyloxyloea leaf extract group were not different from that of the blank control group. The determination data of the extract combination group was not only significantly (P<0.05) higher than that of the blank control group, but also significantly (P<0.05) higher than that of each experimental group in which the extracts were used alone, indicating that the combination of the Dendrobium huoshanense protocorm extract, the Agaricus bisporus extract and the Radermachera xyloxyloea leaf extract had a synergistic effect of improving the NK cell activity of mice.

[0071] The above only describes the preferred embodiments of the present application. It should be noted that the content of the present application can be improved and modified without departing from the technical principles of the present application, and these improvements and modifications should also be considered as the protection scope of the present application.

Claims

1. A functional food product made from the protocorms of Dendrobium huoshanense that enhances immunity, characterized in that, This functional food is composed of 17-19 parts by weight of Dendrobium huoshanense protocorm extract, 11-13 parts by weight of Agaricus bisporus extract, and 7-9 parts by weight of Tofu leaf extract. The preparation method of Dendrobium huoshanense protocorm extract includes the following steps: Preparation of S11 Dendrobium huoshanense protocorm ethanol extract: Dendrobium huoshanense protocorm ultrafine powder was used as raw material and anhydrous ethanol was used as extraction solvent. Under the conditions of vacuum degree 0.1 MPa, ultrasonic power 300 W, temperature 30℃ and material-liquid ratio 1:30, vacuum combined with ultrasonic extraction was performed for 30 min. The extract was concentrated and dried to obtain Dendrobium huoshanense protocorm ethanol extract. Preparation of S12 Dendrobium huoshanense protocorm water extract: using defatted Dendrobium huoshanense protocorm ultrafine powder as raw material and pure water as extraction solvent, vacuum combined with ultrasonic extraction for 30 min under the conditions of vacuum degree 0.1 MPa, ultrasonic power 300 W, temperature 60℃, and material-liquid ratio 1:30, the extract was concentrated and dried to obtain Dendrobium huoshanense protocorm water extract; Preparation of S13 Dendrobium huoshanense protocorm extract: The ethanol extract and water extract of Dendrobium huoshanense protocorm were reconstituted and mixed in equal proportions according to the extraction rate to obtain Dendrobium huoshanense protocorm extract; The preparation method of Agaricus bisporus extract includes the following steps: Preparation of S21 button mushroom ethanol extract: Using button mushroom ultrafine powder as raw material and anhydrous ethanol as extraction solvent, vacuum combined with ultrasonic extraction for 30 min was carried out under the conditions of vacuum degree 0.1 MPa, ultrasonic power 300 W, temperature 30℃, and material-liquid ratio 1:

30. The extract was concentrated and dried to obtain button mushroom ethanol extract. Preparation of S22 Agaricus bisporus water extract: using defatted Agaricus bisporus ultrafine powder as raw material and pure water as extractant, vacuum combined with ultrasonic extraction for 30 min under the conditions of vacuum degree 0.1 MPa, ultrasonic power 300 W, temperature 70℃ and material-liquid ratio 1:30, the extract was concentrated and dried to obtain Agaricus bisporus water extract. Preparation of S23 button mushroom extract: The ethanol extract and water extract of button mushroom were reconstituted and mixed in equal proportions according to the extraction rate to obtain button mushroom extract. The preparation method of *Tofu-shaped Leaf Extract* includes the following steps: Preparation of S31 Tofu Leaf Alcohol Extract: Tofu leaf ultrafine powder was used as raw material and anhydrous ethanol was used as extraction solvent. Under the conditions of vacuum degree 0.1Mpa, ultrasonic power 300W, temperature 30℃ and material-liquid ratio 1:30, vacuum combined with ultrasonic extraction was performed for 30min. The extract was concentrated and dried to obtain the Tofu Leaf Alcohol Extract. Preparation of S32 Tofu Leaf Aqueous Extract: Using defatted tofu leaf ultrafine powder as raw material and pure water as extraction solvent, vacuum combined with ultrasonic extraction for 30 min was performed under the conditions of vacuum degree 0.1 MPa, ultrasonic power 300 W, temperature 80℃, and material-liquid ratio 1:

30. The extract was then concentrated and dried to obtain the tofu leaf aqueous extract. Preparation of S33 Tofu Tree Leaf Extract: The alcohol extract and water extract of Tofu Tree leaves were reconstituted and mixed in equal proportions according to the extraction rate to obtain the Tofu Tree Leaf Extract.